CTF/NF1 transcription factors act as potent genetic insulators for integrating gene transfer vectors
| Autor: | A. Gaussin, Axel Schambach, Christopher Baum, Nicolas Mermod, C. Duros, Ute Modlich, A. Artus, C Bauche, O Cohen-Haguenauer, N J Niederländer |
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| Rok vydání: | 2011 |
| Předmět: |
Transcriptional Activation
CCCTC-Binding Factor Virus Integration Transgene Genetic Vectors Bone Marrow Cells Biology Transfection Viral vector Mice Proto-Oncogenes Genetics Animals Humans Gene silencing Gene Silencing Transgenes RNA Small Interfering Enhancer Molecular Biology Gene Transcription factor Binding Sites Gene Transfer Techniques Provirus MDS1 and EVI1 Complex Locus Protein Friend murine leukemia virus DNA-Binding Proteins Mice Inbred C57BL Repressor Proteins NFI Transcription Factors Enhancer Elements Genetic CTCF Molecular Medicine Insulator Elements HeLa Cells Plasmids Transcription Factors |
| Zdroj: | Gene Therapy. 19:15-24 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2011.70 |
| Popis: | Gene transfer-based therapeutic approaches have greatly benefited from the ability of some viral vectors to efficiently integrate within the cell genome and ensure persistent transmission of newly acquired transgenes to the target cell progeny. However, integration of provirus has been associated with epigenetic repercussions that may influence the expression of both the transgene and cellular genes close to vector integration loci. The exploitation of genetic insulator elements may overcome both issues through their ability to act as barriers that limit transgene silencing and/or as enhancer-blockers preventing the activation of endogenous genes by the vector enhancer. We established quantitative plasmid-based assay systems to screen enhancer-blocker and barrier genetic elements. Short synthetic insulators that bind to nuclear factor-I protein family transcription factors were identified to exert both enhancer-blocker and barrier functions, and were compared to binding sites for the insulator protein CTCF (CCCTC-binding factor). Gamma-retroviral vectors enclosing these insulator elements were produced at titers similar to their non-insulated counterparts and proved to be less genotoxic in an in vitro immortalization assay, yielding lower activation of Evi1 oncogene expression and reduced clonal expansion of bone marrow cells. |
| Databáze: | OpenAIRE |
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